title: A functional glycoprotein competitive recognition and signal amplification strategy for carbohydrate-protein interaction profiling and cell surface carbohydrate expression evaluation.
abstract: A simple and sensitive carbohydrate biosensor has been suggested as a potential tool for accurate analysis of cell surface carbohydrate expression as well as carbohydrate-based therapeutics for a variety of diseases and infections. In this work, a sensitive biosensor for carbohydrate-lectin profiling and in situ cell surface carbohydrate expression was designed by taking advantage of a functional glycoprotein of glucose oxidase acting as both a multivalent recognition unit and a signal amplification probe. Combining the gold nanoparticle catalyzed luminol electrogenerated chemiluminescence and nanocarrier for active biomolecules, the number of cell surface carbohydrate groups could be conveniently read out. The apparent dissociation constant between GOx@Au probes and Con A was detected to be 1.64 nM and was approximately 5 orders of magnitude smaller than that of mannose and Con A, which would arise from the multivalent effect between the probe and Con A. Both glycoproteins and gold nanoparticles contribute to the high affinity between carbohydrates and lectin. The as-proposed biosensor exhibits excellent analytical performance towards the cytosensing of K562 cells with a detection limit of 18 cells, and the mannose moieties on a single K562 cell were determined to be 1.8 × 10(10). The biosensor can also act as a useful tool for antibacterial drug screening and mechanism investigation. This strategy integrates the excellent biocompatibility and multivalent recognition of glycoproteins as well as the significant enzymatic catalysis and gold nanoparticle signal amplification, and avoids the cell pretreatment and labelling process. This would contribute to the glycomic analysis and the understanding of complex native glycan-related biological processes.
authors.affiliation: Department of Chemistry, Beijing Key Laboratory for Microanalytical Methods and Instrumentation, Tsinghua University, Beijing 100084, China.
authors.forename: Yangzhong
authors.initials: Y
authors.lastname: Wang
authors.forename: Zhuhai
authors.initials: Z
authors.lastname: Chen
authors.forename: Yang
authors.initials: Y
authors.lastname: Liu
authors.forename: Jinghong
authors.initials: J
authors.lastname: Li
mesh_terms: D055162:Biocatalysis; D015374:Biosensing Techniques; D002241:Carbohydrates / Q000737:chemistry*; D002462:Cell Membrane / Q000737:chemistry / Q000378:metabolism; D003208:Concanavalin A / Q000378:metabolism; D055664:Electrochemical Techniques; D004566:Electrodes; D004800:Enzymes, Immobilized / Q000737:chemistry / Q000378:metabolism; D005949:Glucose Oxidase / Q000737:chemistry / Q000378:metabolism; D006023:Glycoproteins / Q000737:chemistry / Q000378:metabolism*; D006046:Gold / Q000737:chemistry; D006801:Humans; D020014:K562 Cells; D037102:Lectins / Q000737:chemistry / Q000378:metabolism; D008163:Luminescent Measurements; D053768:Metal Nanoparticles / Q000737:chemistry
chemical_list: D002241:Carbohydrates; D004800:Enzymes, Immobilized; D006023:Glycoproteins; D037102:Lectins; D003208:Concanavalin A; D006046:Gold; D005949:Glucose Oxidase